Study On Plant Regeneration System And The Influence Factors Genetic Transformation Of Xinjiang Cotton Varieties In Vitro

Cotton is an important economical crop all over the world. Requirement for cotton varities'characters is complicated, because phases of its growth and development nicely was consistent with seasons of high frequency of various natural disaster. The conventional breeding proposal is faced with spareness of germ plasm resource and trouble of remote ancestors'crosses, which make good characters'combination be greatly inconvenience, especially traits of high yield, good quality and muti-resistance to diseases and insects. So more and more scientific researchers pay attention to the study on improving cotton by genetic engineering technology. But many obstacles still exist in development of cotton biology at present, for example somatic embryogenesis of cotton is genetype-dependent, tissue culture of cotton is low efficiency (take time and energy), genetic transformation technology is intricate (repeatness and transformation are low), and variation of transgenic cotton's agricultural traits is complex, and so on. These questions have become hindrance and choke point of cotton biology, validatation of cotton function gene and cotton relative gene clone.As the problems narrated above, The paper deal with four varieties of Xinjiang cotton, Study on high efficiency plant regeneration system and the influence factors genetic transformation of Xinjiang Cotton Varieties in vitro.The primary research results are as follows.1 The research deal with four varieties of Xinjiang cotton, epicotyls act as the explants in vitro.The explants were cultured on four different shoot inducement culture mediums. In the culture medium of MSB + 6-BA 0.3mg/L + NAA 0.01mg/L had the highest ratio of producing multiple shoots. But when the concentration of 6-BA is increased,the vitrification phenomenon is pricked up.2 The research deal with four varieties of Xinjiang cotton, epicotyls act as the explants in vitro.The explants were cultured on four different shoot inducement culture mediums.Among the different shoot inducement culture mediums, In the culture medium of MSB + KT 0.05mg/L + NAA 0.01mg/L had the highest ratio of producing multiple shoots.But the ratio of producing multiple shoots are all lower than the compounding of 6-BA and NAA.The growth instance of multiple shoots is disproportion.3 Rooting was induced in 1/4MS added 10 mg/L IBA is the best than any other rooting culture mediums,and its ratio can reach 86.7% Shoot induction of cotton epicotyl is genetype-independent, the results were not appeared much difference between these cultivars. It is showed that multiple shoots induction was not under the control of different genotype.The plant regeneration system is high efficiency, short cycle and maneuverable.4 The agrobacterium-mediated gene transformation procedure was discussed in detail. The factors, including precultivation time, immersion time, cocultivation periods and concentration of acetosyringone (AS), The optimized transformation protocol was obtained, when epicotyl explants precultured for 3~5 days on regeneration medium were immersed into the diluted agrobacterium suspension for 15 min, and then were co-cultured containing 100mg/L AS for 2 days with agrobacterium,and were finally subcultured on selection medium after reinforcing cultivation for 3 days on the regeneration medium containing 800 mg/L cef and 50mg/L Kan.5 From treated epicotyls by agrobacterium-mediated transformation,resistant plantlets have been obtained after kan resistance screening.Under irradiation of blue fluorescence, the leaf of resistant plantlets were primary inspected,the green fluorescence could be observed from the leaf of resistant plantlets.